The Toll-like receptors (TLRs) are a family of pattern recognition receptors (PRRs) that recognize a diverse set of Pathogen Associated Molecular Patterns (PAMPs) from different classes of microbes and play an essential role in innate immune defense of the host. In addition, TLRs have been shown to play a critical role in induction of adaptive immune responses. Recognition of PAMPs by TLRs on dendritic cells (DCs) leads to DC maturation, a process that involves enhanced expression of MHC and co-stimulatory molecules as well as migration to the draining lymph nodes to prime naive T cells. TLR activation also leads to production of pro-inflammatory cytokines that contribute to CD4 T cell activation by overcoming Treg mediated suppression. Notably, TLR expression is not limited to DCs. In addition to cells of myeloid origin such as DCs and macrophages, other cells such as neutrophils, epithelial cells, fibroblasts, B lymphocytes and T lymphocytes have been shown to express functional TLRs. Although the outcome of TLR activation in all of these cell types is quite different, their individual contribution to the activation of naive CD4 T cells is not well understood. The overall goals of this study are to characterize TLR dependent mechanisms that control activation of naive CD4 T cells and their differentiation into long lived memory cells. Three specific aims are proposed to study the role of TLRs in controlling adaptive immunity. Aim 1 will characterize the role of different TLR expressing cells in naive CD4 T cell activation. Aim 2 will analyze the contribution of cytokines secreted by DCs, particularly Interleukin-1 and Interleukin-6, to CD4 T cell activation in vivo. Aim 3 proposes to understand the mechanisms by which TLR activation of DCs contributes to generation of CD4 T cell memory. Together these studies will yield novel insights into how recognition of pathogens via TLRs shapes CD4 T cell responses and will lead to better understanding of the mechanisms of innate control of adaptive immune responses in vivo. Furthermore, this work will aid in designing effective vaccine formulations for prophylactic vaccinations. PUBLIC HEALTH RELEVANCE: Humans and other multicellular organisms are exposed to a wide variety of different pathogens. The immune system has specialized receptors called Toll-like receptors (TLRs) to recognize structural and metabolic components of pathogenic microbes. This recognition is important for immediate defense of the host by the innate immune system. Long-term protection from re-infection however requires activation of pathogen specific T cells. T cell activation has been shown to be dependent on prior activation of TLRs on the cells of the innate immune system, namely dendritic cells. In this proposal, we will study the mechanisms by which TLR activation in dendritic cells contributes to induction of long- term protective immunity by T cells. The knowledge acquired from these studies will aid in development of new vaccination strategies against pathogens